Today I checked the T2 lines of wild type CYP83B135S:CDS:YFP:HA in Ws-2 and gul4 background in Basta MS plates, I selected the 3 lines from severe phenotype 2 from moderate and 2 from weak phenotype, surpizingly the T2  liness in gul4 background didn't show the complete wild type phenotype despite of their WT phenotype in T1 in gul4 background. All the seedlings  show  a big opened  leaves more similar to WT but regarding the hypocotyl some of them show wild type but the others show gul4 like long hypocotyl. today I will try to count the no. of sedling showing different phenotype and will collect  the tissue for western to confirm the translation of transgene is ok.

Yesterday,

1. I continued the western blot, only today I can get the results.

2. Seed harvesting of F1of gul4 and atr4-1 and atr4-2.

3. LR cloning of gul4 OX lines using competent cell from Prof. Noh's lab since last time LR I could not get any colony. I am not sure but may the problem is with competent cell. The competent cell, DH5a, in regular use is out of so I use the XL blue found in -80 freezer, and not mentioned when was it prepared and who prepared.

 Today,

1. Completion of Western blot.

2. Counting of seedling with different phenotype in T2  of CYP83B1 OX lines in gul4 background and tissue collection from OX lines in WT and gul4 background .

3. sequencing results analysis of entry of gul4 promoter.

4. Tissue collection of srd from srd mapping population.

5. GUS staining of  DWF4pro:GUS in gul4 background  selecting the KAn resistant and gul4 phenotype seedling from F2 of gul4X p6-1 in Kan MS.